一株全基因合成的高温嗜碱甘露聚糖酶的酶学性质分析

Analysis Enzymatic Characterization of A Gene Synthesis of Thermostable and Alkaliphiles Endo-β-1,4-mannanase

根据来源于褐色高温单孢菌(Thermomonospora fusca)YX基因组序列,预测并合成了甘露聚糖酶ManB全基因序列(Accession No.KJ806638),与表达载体pPIC9K重组后,转化毕赤酵母(Pichia pastoris)GS115,筛选获得重组菌株ManB-GS115。其酶学性质检测结果表明,该酶最适温度为70℃,最适pH为9.0。该甘露聚糖酶ManB在碱性高温环境下有较高的酶活力,可应用于食品、酿造、饲料、纺织和医药等工业领域。

According to the known genomic sequences of Thermomonospora fusca YX and the codon bias of yeast,the opti- mized mannanase ManB sequence （Accession No. KJ806638） was designed and synthesized. The ManB gene was cloned into the pPIC9K vector,and then expressed in Pichia pastoris GSll5 to obtain ManB. The enzymatic properties were shown that the optimal reaction temperature was 70 ℃ and pH was 9.0. With the excellent enzymatic properties,the mannanase ManB could be potentially applied in brewing, food, feed, textileand pharmaceuticals fields.